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Toxicity bioassay of waste cooking oil-based biodiesel on marine microalgae.

ABSTRACT: The world biodiesel production is increasing at a rapid rate. Despite its perceived safety for the environment, more detailed toxicity studies are mandatory, especially in the field of aquatic toxicology. While considerable attention has been paid to biodiesel combustion emissions, the toxicity of biodiesel in the aquatic environment has been poorly understood. In our study, we used an algae culture growth-inhibition test (OECD 201) for the comparison of the toxicity of B100 (pure biodiesel), produced by methanol transesterification of waste cooking oil (yellow grease), B0 (petroleum diesel fuel) and B20 (diesel-biodiesel blended of 20% biodiesel and 80% petroleum diesel fuel by volume). Two marine diatoms Attheya ussuriensis and Chaetoceros muelleri, the red algae Porphyridium purpureum and Raphidophyte Heterosigma akashiwo were employed as the aquatic test organisms. A sample of biodiesel from waste cooking oil without dilution with petroleum diesel (B100) showed the highest level of toxicity for the microalgae A. ussuriensis, C. muelleri and H. akashiwo, compared to hexane, methanol, petroleum diesel (B0) and diluted sample (B20). The acute EC₅₀ in the growth-inhibition test (96 h exposure) of B100 for the four species was in the range of 3.75-23.95 g/L whereas the chronic toxicity EC₅₀ (7d exposure) was in the range of 0.42-16.09 g/L.

SUBMITTER: Pikula KS

PROVIDER: S-EPMC6317304 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Oleaginous yeasts are fast emerging as a possible feedstock for biodiesel production. Yarrowia lipolytica, a model oleaginous yeast is known to utilize a variety of hydrophobic substrates for lipid accumulation including waste cooking oil (WCO). Approaches to increase lipid content in this yeast include metabolic engineering which requires manipulation of multiple genes in the lipid biosynthesis pathway. A classical and cost-effective approach, namely, random chemical mutagenesis on the yeast can lead to increased production of biodiesel as is explored here.In this study, chemical mutagenesis using the alkylating agent, N- methyl-N'-nitro-N-nitrosoguanidine (MNNG) as well as an additional treatment with cerulenin, a fatty acid synthase inhibitor generated 800 mutants of Y. lipolytica NCIM 3589 (761 MNNG treated and 39 MNNG + cerulenin treated). A three-stage screening using Sudan Black B plate technique, Nile red fluorimetry and total lipid extraction using solvent was performed, which enabled selection of ten high lipid yielding mutants. Time course studies of all the ten mutants were further undertaken in terms of biomass, lipid yield and lipid content to select three stable mutants (YlB6, YlC7 and YlE1) capable of growing and accumulating lipid on WCO, with lipid contents of 55, 60 and 67% as compared to 45% for the wild type. The mutants demonstrated increased volumetric lipid productivities (0.062, 0.044 and 0.041 g L-1 h-1) as compared to the wild type (0.033 g L-1 h-1). The fatty acid profile of the three mutants consisted of a high content of C16 and C18 saturated and monounsaturated fatty acids and was found to be suitable for biodiesel production. The fuel properties, namely, density, kinematic viscosity, total acid number, iodine value of the three mutants were evaluated and found to lie within the limits specified by internationally accepted standards. Additionally, it was noted that the mutants demonstrated better cetane numbers and higher heating values than the wild type strain.The chemical mutagenesis strategy adopted in this study resulted in the successful isolation of three stable high SCO yielding mutants. The mutants, namely, YlB6, YlC7 and YlE1 exhibited a 1.22, 1.33 and 1.49-fold increase in lipid contents when grown on 100 g L-1 waste cooking oil than the parental yeast strain. The fatty acid methyl ester (FAME) profiles of all the three mutants was determined to be suitable for biodiesel suggesting their potential applicability while simultaneously addressing the management of waste cooking oil.

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Toxicity bioassay of waste cooking oil-based biodiesel on marine microalgae.

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